Fluorescence Microscopy



To harness the power of fluorescence for biological microscopic imaging, a number of additional components must be introduced to the standard light microscope. The most important modifications are (1) a strong illumination system that provides suitable wavelengths for exciting a specific fluorochrome, (2) some mechanism to limit the illumination beam to specific wavelengths so that only the fluorochrome of choice is excited, and (3) a means to image only the light emitted from the fluorochrome so that the excitation light (and other stray wavelengths) does not degrade the image. In this chapter, we will discuss a basic set-up that meets these criteria. Subsequent chapters will expand on this theme and discuss additional modifications required for laser scanning and spinning-disk confocal fluorescence.


Filters Fluorochrome Laser Magnification Objective lens Refractive index Refractive index mismatch 


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of PathologyVanderbilt University Medical CenterSouth NashvilleUSA

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